Cargando…

Characterisation of SARS-CoV-2 Lentiviral Pseudotypes and Correlation between Pseudotype-Based Neutralisation Assays and Live Virus-Based Micro Neutralisation Assays

The recent outbreak of a novel Coronavirus (SARS-CoV-2) and its rapid spread across the continents has generated an urgent need for assays to detect the neutralising activity of human sera or human monoclonal antibodies against SARS-CoV-2 spike protein and to evaluate the serological immunity in hum...

Descripción completa

Detalles Bibliográficos
Autores principales: Hyseni, Inesa, Molesti, Eleonora, Benincasa, Linda, Piu, Pietro, Casa, Elisa, Temperton, Nigel J, Manenti, Alessandro, Montomoli, Emanuele
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7551040/
https://www.ncbi.nlm.nih.gov/pubmed/32927639
http://dx.doi.org/10.3390/v12091011
_version_ 1783593095104823296
author Hyseni, Inesa
Molesti, Eleonora
Benincasa, Linda
Piu, Pietro
Casa, Elisa
Temperton, Nigel J
Manenti, Alessandro
Montomoli, Emanuele
author_facet Hyseni, Inesa
Molesti, Eleonora
Benincasa, Linda
Piu, Pietro
Casa, Elisa
Temperton, Nigel J
Manenti, Alessandro
Montomoli, Emanuele
author_sort Hyseni, Inesa
collection PubMed
description The recent outbreak of a novel Coronavirus (SARS-CoV-2) and its rapid spread across the continents has generated an urgent need for assays to detect the neutralising activity of human sera or human monoclonal antibodies against SARS-CoV-2 spike protein and to evaluate the serological immunity in humans. Since the accessibility of live virus microneutralisation (MN) assays with SARS-CoV-2 is limited and requires enhanced bio-containment, the approach based on “pseudotyping” can be considered a useful complement to other serological assays. After fully characterising lentiviral pseudotypes bearing the SARS-CoV-2 spike protein, we employed them in pseudotype-based neutralisation assays in order to profile the neutralising activity of human serum samples from an Italian sero-epidemiological study. The results obtained with pseudotype-based neutralisation assays mirrored those obtained when the same panel of sera was tested against the wild type virus, showing an evident convergence of the pseudotype-based neutralisation and MN results. The overall results lead to the conclusion that the pseudotype-based neutralisation assay is a valid alternative to using the wild-type strain, and although this system needs to be optimised and standardised, it can not only complement the classical serological methods, but also allows serological assessments to be made when other methods cannot be employed, especially in a human pandemic context.
format Online
Article
Text
id pubmed-7551040
institution National Center for Biotechnology Information
language English
publishDate 2020
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-75510402020-10-16 Characterisation of SARS-CoV-2 Lentiviral Pseudotypes and Correlation between Pseudotype-Based Neutralisation Assays and Live Virus-Based Micro Neutralisation Assays Hyseni, Inesa Molesti, Eleonora Benincasa, Linda Piu, Pietro Casa, Elisa Temperton, Nigel J Manenti, Alessandro Montomoli, Emanuele Viruses Article The recent outbreak of a novel Coronavirus (SARS-CoV-2) and its rapid spread across the continents has generated an urgent need for assays to detect the neutralising activity of human sera or human monoclonal antibodies against SARS-CoV-2 spike protein and to evaluate the serological immunity in humans. Since the accessibility of live virus microneutralisation (MN) assays with SARS-CoV-2 is limited and requires enhanced bio-containment, the approach based on “pseudotyping” can be considered a useful complement to other serological assays. After fully characterising lentiviral pseudotypes bearing the SARS-CoV-2 spike protein, we employed them in pseudotype-based neutralisation assays in order to profile the neutralising activity of human serum samples from an Italian sero-epidemiological study. The results obtained with pseudotype-based neutralisation assays mirrored those obtained when the same panel of sera was tested against the wild type virus, showing an evident convergence of the pseudotype-based neutralisation and MN results. The overall results lead to the conclusion that the pseudotype-based neutralisation assay is a valid alternative to using the wild-type strain, and although this system needs to be optimised and standardised, it can not only complement the classical serological methods, but also allows serological assessments to be made when other methods cannot be employed, especially in a human pandemic context. MDPI 2020-09-10 /pmc/articles/PMC7551040/ /pubmed/32927639 http://dx.doi.org/10.3390/v12091011 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Hyseni, Inesa
Molesti, Eleonora
Benincasa, Linda
Piu, Pietro
Casa, Elisa
Temperton, Nigel J
Manenti, Alessandro
Montomoli, Emanuele
Characterisation of SARS-CoV-2 Lentiviral Pseudotypes and Correlation between Pseudotype-Based Neutralisation Assays and Live Virus-Based Micro Neutralisation Assays
title Characterisation of SARS-CoV-2 Lentiviral Pseudotypes and Correlation between Pseudotype-Based Neutralisation Assays and Live Virus-Based Micro Neutralisation Assays
title_full Characterisation of SARS-CoV-2 Lentiviral Pseudotypes and Correlation between Pseudotype-Based Neutralisation Assays and Live Virus-Based Micro Neutralisation Assays
title_fullStr Characterisation of SARS-CoV-2 Lentiviral Pseudotypes and Correlation between Pseudotype-Based Neutralisation Assays and Live Virus-Based Micro Neutralisation Assays
title_full_unstemmed Characterisation of SARS-CoV-2 Lentiviral Pseudotypes and Correlation between Pseudotype-Based Neutralisation Assays and Live Virus-Based Micro Neutralisation Assays
title_short Characterisation of SARS-CoV-2 Lentiviral Pseudotypes and Correlation between Pseudotype-Based Neutralisation Assays and Live Virus-Based Micro Neutralisation Assays
title_sort characterisation of sars-cov-2 lentiviral pseudotypes and correlation between pseudotype-based neutralisation assays and live virus-based micro neutralisation assays
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7551040/
https://www.ncbi.nlm.nih.gov/pubmed/32927639
http://dx.doi.org/10.3390/v12091011
work_keys_str_mv AT hyseniinesa characterisationofsarscov2lentiviralpseudotypesandcorrelationbetweenpseudotypebasedneutralisationassaysandlivevirusbasedmicroneutralisationassays
AT molestieleonora characterisationofsarscov2lentiviralpseudotypesandcorrelationbetweenpseudotypebasedneutralisationassaysandlivevirusbasedmicroneutralisationassays
AT benincasalinda characterisationofsarscov2lentiviralpseudotypesandcorrelationbetweenpseudotypebasedneutralisationassaysandlivevirusbasedmicroneutralisationassays
AT piupietro characterisationofsarscov2lentiviralpseudotypesandcorrelationbetweenpseudotypebasedneutralisationassaysandlivevirusbasedmicroneutralisationassays
AT casaelisa characterisationofsarscov2lentiviralpseudotypesandcorrelationbetweenpseudotypebasedneutralisationassaysandlivevirusbasedmicroneutralisationassays
AT tempertonnigelj characterisationofsarscov2lentiviralpseudotypesandcorrelationbetweenpseudotypebasedneutralisationassaysandlivevirusbasedmicroneutralisationassays
AT manentialessandro characterisationofsarscov2lentiviralpseudotypesandcorrelationbetweenpseudotypebasedneutralisationassaysandlivevirusbasedmicroneutralisationassays
AT montomoliemanuele characterisationofsarscov2lentiviralpseudotypesandcorrelationbetweenpseudotypebasedneutralisationassaysandlivevirusbasedmicroneutralisationassays